Schwartz B, Hittelman A, Daneshvar L, Basu H S, Marton L J, Feuerstein B G
Department of Neurological Surgery, School of Medicine, University of California, San Francisco 94143, USA.
Biochem J. 1995 Nov 15;312 ( Pt 1)(Pt 1):83-90. doi: 10.1042/bj3120083.
Ornithine decarboxylase (ODC) is a rate-determining enzyme of the polyamine-biosynthetic pathway. We sought to produce cells with impaired ODC function in order to study the biological functions of polyamines. Saccharomyces cerevisiae strains were obtained by one-step gene replacement of a 900 bp fragment of the yeast ODC gene (SPE1) with the yeast URA3 gene. Spores derived from SPE1/spe1 cells germinated at reduced efficiency relative to SPE1/SPE1. Sustained growth of spe1 haploid mutants in polyamine-free medium led to intracellular polyamine depletion, reduction in budding index, G1 arrest and cessation of growth, and cells that were large and misshapen. All of these effects were completely reversed by adding polyamines to the medium, even after 5 days of polyamine starvation. A diploid yeast strain bearing two copies of disrupted spe1 lost heterozygosity at the mating-type locus more often when grown in the absence of polyamines than when grown in their presence, indicating that polyamine deficiency leads to either chromosome loss or to mitotic recombination.
鸟氨酸脱羧酶(ODC)是多胺生物合成途径中的一种限速酶。我们试图构建ODC功能受损的细胞,以便研究多胺的生物学功能。通过用酵母URA3基因一步置换酵母ODC基因(SPE1)的900 bp片段,获得了酿酒酵母菌株。相对于SPE1/SPE1,来自SPE1/spe1细胞的孢子萌发效率降低。spe1单倍体突变体在无多胺培养基中的持续生长导致细胞内多胺耗竭、出芽指数降低、G1期阻滞和生长停止,细胞变得又大又畸形。即使在多胺饥饿5天后,向培养基中添加多胺也能完全逆转所有这些效应。与在有polyamines存在的情况下生长相比,携带两个 disrupted spe1 拷贝的二倍体酵母菌株在没有 polyamines 的情况下生长时,在交配型位点更频繁地失去杂合性,这表明多胺缺乏会导致染色体丢失或有丝分裂重组。